The major objectives of the proposed research are fivefold: 1) The isolation, purification, and study of the mechanism of reaction of two alpha-fucosyltransferases, three beta-galactosyltransferases, one alpha-galactosyltransferase, one alpha-sialytransferase and one beta-glucosyltransferase which are involved in the biosynthesis of neutral (globoside and blood group family) and acidic (ganglioside family) glycosphingolipids. The subcellular distribution of these enzymes in normal tissues will be determined. Once the methods have been established, further studies of these enzyme levels in human normal and pathological tissues (Tay-Sachs, multiple sclerosis, etc.) will be made. 2) Structure determination and immunological characterization of a pentaglycosylceramide isolated from bovine erythrocytes will be completed. The structure of branched chain glycosphingolipids in bovine and ovine erythrocytes will be investigated. 3) Biosynthetic studies in vitro of blood group H(I and II), B(I and II), A(I and II), novel Lea, sialyl novel Lea, and fucosyl GM1 ganglioside in bone marrow, serum, spleen, testis, and kidney of humans, rabbits, cattle, sheep, goats, hogs, and guinea pigs will be continued and extended. Attempts will be made to determine the presence of fucose-containing blood group type glycosphingolipids on eukaryotic higher primate cell surfaces (e.g., African green monkey kidney, Vero, and human lung, WI-38, cells). Their possible role in cellular interaction and binding to specific plant lectins (e.g., Ulex europeus, Dolichos biflorus, and Bandeiraea simplicifolia) will be investigated. 4) Isolation, characterization, and purification of an alpha-fucosidase from clam (Venus mercenaria) liver and a beta-galactosidase from green papaya will be continued. These enzymes are useful for anomeric structure determination of the biosynthetic glycosphingolipids. 5) Metabolic changes in neuronal cells (NS-20, N1E-115, N-18, and IMR-32) will be studied before and after chemically induced differentiation.